LOCUS       sequence-297408-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6753)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4252956
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             119..139
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             140..217
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    286..407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(297..316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     351..370
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      522..568
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      598..734
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    779..850
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1076..1664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1156..1175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2164..3373
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3210..3229
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3377..4405
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4432..4480
                     /note="synthetic polyadenylation signal"
     CDS             join(4746..4814,4815..5606)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4964..4983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5619..5755
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5907..5978
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5983..6029)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6047..6095
                     /note="synthetic polyadenylation signal"
     enhancer        6176..6479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6481..6683
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6634..6654
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6752..6753,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggg tacataaatg ttttcgcctt tccattcacg atctattcac tccttttatg
       61 taggatgaac tctcgcaatt acattgctca ggtagacgtc gtgaacttta acctaactga
      121 aaacctttac tttcagtcga agaggcgatg gaaaaagaat tttatcgcgg tgagtgcagc
      181 caacagattc aaaaagatat cctcgtctgg tgccctctaa ggatccgcag gcctctgcta
      241 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
      301 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      361 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      421 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      481 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
      541 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
      601 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
      661 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
      721 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
      781 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
      841 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
      901 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
      961 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1021 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1081 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1141 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1201 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1261 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1321 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1381 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1441 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1501 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1561 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1621 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1681 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1741 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1801 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1861 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     1921 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     1981 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     2041 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     2101 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     2161 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     2221 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     2281 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2341 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     2401 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     2461 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     2521 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     2581 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     2641 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     2701 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     2761 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     2821 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     2881 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     2941 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3001 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt
     3061 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     3121 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     3181 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     3241 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     3301 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     3361 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3421 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3481 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3541 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     3601 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     3661 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3721 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     3781 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     3841 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     3901 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     3961 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4021 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4081 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4141 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4201 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4261 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4321 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4381 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     4441 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     4501 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     4561 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     4621 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     4681 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     4741 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4801 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4861 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4921 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4981 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5041 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5101 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5161 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5221 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5281 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     5341 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     5401 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5461 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5521 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5581 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     5641 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     5701 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     5761 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     5821 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     5881 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     5941 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     6001 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     6061 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     6121 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     6481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6661 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     6721 taccatccac tcgacacacc cgccagcggc cgc
//
